Abstract
A convenient technology to quantify three-dimensional (3D) morphological features would have widespread applications in biomedical research. Based on combined improvements in sample preparation, tomographic imaging and computational processing, we present a procedure for high-resolution 3D quantification of structures within intact adult mouse organs. Using the nonobese diabetic (NOD) mouse model, we demonstrate a correlation between total islet β-cell volume and the onset of type-1 diabetes.
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Acknowledgements
We thank T. Bock and B. Hellman for helpful discussions. This work was supported by the Medical Research Council (H.M, J.S), the Wenner-Gren foundation (C.E.L), Novo Nordisk Fonden, the Swedish Diabetes Association (D.H), the Juvenile Diabetes Research Foundation (D.H, J.S, U.A), the Swedish Research Council (D.H, U.A) and the Kempe Foundation (U.A.).
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Contributions
T.A. developed the immunohistochemical protocol and performed immmunohistochemical analyses; A.A. performed the OPT and confocal scanning; H.M. performed OPT reconstructions and islet quantifications; C.E.L performed histological integrity controls; D.H. designed the NOD experimental setup; J.S. developed the subtraction script and supervised the project; U.A. initiated and supervised the project.
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J.S. is the inventor on four patents relating to OPT technology.
Supplementary information
Supplementary Fig. 1
Optimized labeling allows visualization of large-scale specimen.
Supplementary Fig. 2
Example of variation in islet composition for a non-diabetic versus an overtly diabetic NOD mouse.
Supplementary Fig. 3
3D reconstructions of an intact mouse brain labeled for neuronal marker Isl1.
Supplementary Video 1
Iso-surface reconstruction of an adult wild type splenic pancreas. Movie showing a 3D OPT image of an intact splenic mouse pancreas (8 weeks). The reconstruction of the pancreas outline is based on the autofluorescence signal (grey) whereas the insulin expressing cells of the islets (red) are based on the signal from insulin specific antibodies. The specimen is ∼ 1.2 × 0.7 cm.
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Alanentalo, T., Asayesh, A., Morrison, H. et al. Tomographic molecular imaging and 3D quantification within adult mouse organs. Nat Methods 4, 31–33 (2007). https://doi.org/10.1038/nmeth985
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DOI: https://doi.org/10.1038/nmeth985
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